Abstract
In recent years, one of the major breakthroughs in eukaryotic biology has been the discovery of the RNA interference (RNAi) pathway [1], a powerful gene silencing mechanism that operates both at the transcriptional and post-transcriptional levels. Originally discovered as a mechanism through which double-stranded RNA (dsRNA) triggers degradation of homologous transcripts, the RNAi pathway is essential for a variety of gene silencing phenomena [2], including heterochromatin formation, DNA and histone methylation, macronuclear DNA elimination in Tetrahymena, promoter silencing in plants, and translational control by micro RNAs. Furthermore, in certain organisms RNAi appears to provide or has been proposed to provide a genome defense mechanism to limit the spreading of mobile elements [3–9] and in plants it is a powerful antiviral response [10]. At the genetic level there are two proteins that are the universal hallmark of the RNAi pathway: Dicer, an RNAse III-related enzyme, and a member of the Argonaute protein family [2]. In the ‘classical’ RNAi pathway, which is triggered by long dsRNAs, Dicer [11] is the endonuclease that processes dsRNA into 20–30 nt small interfering RNAs (siRNAs), whereas an Argonaute family member provides the endonuclease activity or “Slicer” that cleaves transcripts after base-pairing with complementary siRNAs [12–16].
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